CN114875150A - DUB1 gene, and application of detection reagent and inhibitor thereof - Google Patents

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a DUB1 gene, and an application of a detection reagent and an inhibitor thereof. The present invention demonstrates that the expression of DUB1 gene is elevated in gastric cancer and is associated with low survival rate. By detecting the expression level of the DUB1 gene in a subject, it is possible to judge whether the subject has gastric cancer and the risk of having gastric cancer. The consumption of DUB1 can inhibit the development of gastric cancer in vivo and in vitro, so that the application of DUB1 gene as a target point to treat gastric cancer has better feasibility and popularization.

Description

DUB1 gene, and application of detection reagent and inhibitor thereof

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to a DUB1 gene, and an application of a detection reagent and an inhibitor thereof.

Background

The carcinogenic process of gastric cancer is a pathological process including gastritis, intestinal metaplasia, and epithelial neoplasia. Due to the occult onset and mild symptoms, most cancer patients are found in patients already at a later stage. Its 5-year survival rate is only 35%. Several biomedical studies have shown that H.pylori infection and chronic inflammation are involved in gastric cancer development, but the specific mechanism of gastric cancer progression is not clear. Therefore, the discovery of new gastric cancer biomarkers and therapeutic targets for gastric cancer is urgent and essential.

Several studies have found a potential link between the Hippo signaling pathway and gastric cancer. The Hippo signaling pathway plays an important role in multiple biological processes such as organ size control, tissue homeostasis, carcinogenesis, and immune response. Activation of Hippo signaling is subject to a phosphorylation cascade, mammalian Hippo kinase MST1/2 phosphorylates LATS1/2 kinase, while phosphorylated LATS1/2 promotes phosphorylation of YAP/TAZ proteins, leading to cytoplasmic localization and protein degradation. If the Hippo signaling pathway is inactivated, YAP/TAZ will be unphosphorylated, translocated to the nucleus, and interact with multiple transcription factors, promoting the expression of its target genes. YAP/TAZ overexpression inhibits the progression of gastric cancer in a H.pylori-induced mouse gastric cancer model. However, despite the considerable efforts to develop drugs targeting Hippo signaling, their clinical applications are still in an immature stage.

TAZ (a transcriptional co-activator with PDZ binding motifs) is a downstream effect of Hippo signaling, a transcription co-factor for multiple transcription factors, including TEADs. Several studies have shown that TAZ may be a key factor in the carcinogenesis of human cancers. For example, molecular studies have shown that activation of TAZ can transactivate a variety of signaling pathways, such as the Wnt and Heddehog signaling pathways. In human cancer samples, levels of TAZ protein are elevated in breast and gastric cancers, and expression levels are associated with poor prognosis. Thus, targeting TAZ proteins could be candidate components of the ubiquitin-proteasome system, and could be used to rescue the tumor-suppressor function of the Hippo signaling pathway and the progression of gastric cancer progression.

Regulation of protein stability is influenced by the balance between E3 ubiquitin ligase and deubiquitinase, which is critical to maintaining normal physiological homeostasis. Disruption of protein ubiquitination regulation may lead to a variety of human diseases, including cancer. We and other groups have found that several E3 ubiquitin ligases act on Hippo pathway effectors, modulating Hippo signalling activity and cancer progression. Although several deubiquitinases have been reported to be involved in the regulation of Hippo signaling, it is unclear which deubiquitinase has an important effect.

Disclosure of Invention

In order to make up for the deficiencies of the prior art, the invention particularly provides a DUB1 gene, and related applications of a detection reagent and an inhibitor thereof in diagnosis and treatment of gastric cancer.

In a first aspect, the present invention provides the use of a reagent for detecting the expression level of DUB1 gene in the preparation of a product for diagnosing gastric cancer.

Further, the DUB1 gene is expressed in a patient with gastric cancer.

Still further, the agent is selected from:

a probe that specifically recognizes the DUB1 gene; or

Primers that specifically amplify the DUB1 gene; or

A specific binding agent that specifically binds to a protein encoded by the DUB1 gene.

Furthermore, the primer sequence for specifically amplifying the DUB1 gene is shown in SEQ ID NO. 9-10.

In a second aspect, the invention provides the use of the DUB1 gene in screening potential substances for treating gastric cancer, wherein the substances down-regulate the expression level of the DUB1 gene.

In a third aspect, the invention provides an application of the expression inhibitor of the DUB1 gene in preparation of a medicament for treating gastric cancer.

Further, the inhibitor comprises siRNA capable of targeting and interfering with the DUB1 gene.

Furthermore, the sequence of the siRNA is shown in SEQ ID NO. 3-6.

In a third aspect, the present invention provides a pharmaceutical composition for the treatment of gastric cancer, comprising a therapeutically effective amount of the inhibitor.

Furthermore, the pharmaceutical composition also comprises other medicines compatible with the inhibitor and a pharmaceutically acceptable carrier and/or auxiliary material.

The invention has the beneficial effects that:

according to the invention, the DUB1 gene is found to be related to the occurrence of gastric cancer in clinical samples and experimental studies for the first time. We confirmed that DUB1 gene expression is elevated in gastric cancer and associated with low survival. By detecting the expression level of the DUB1 gene in a subject, it is possible to judge whether the subject has gastric cancer and the risk of having gastric cancer.

The invention discovers for the first time that consumption of DUB1 can inhibit the development of gastric cancer in vivo and in vitro, so that the application of DUB1 gene as a target point to treat gastric cancer has better feasibility and popularization.

Drawings

FIG. 1 is a box plot of the expression levels of DUB1 mRNA in gastric cancer tissues and normal paracancerous tissues at different stages of TCGA data analysis.

FIG. 2 is a graph showing the results of differential expression of DUB1 protein in gastric cancer tissues and normal paracancerous tissues in tissue chips and immunohistochemical analysis.

FIG. 3 shows the silencing efficiency verification of siRNA targeting DUB1 gene, FIG. 3A shows the Western blot detection result of MGC803 cell, FIG. 3B shows the Western blot detection result of AGS cell, FIG. 3C shows the RT-qPCR detection result of MGC803 cell, and FIG. 3D shows the RT-qPCR detection result of AGS cell.

Fig. 4 shows the results of measurement of the effect of siRNA targeting DUB1 gene on gastric cancer cell activity, panel a shows MGC803 cell, and panel B shows AGS cell.

Fig. 5 is a result of measuring the effect of siRNA targeting DUB1 gene on EdU positive cells in gastric cancer cells, panel a is an image analysis result of MGC803 cells, panel B is a detailed quantification result of panel a, panel C is an image analysis result of AGS cells, and panel D is a detailed quantification result of panel C.

Fig. 6 shows the results of measuring the effect of siRNA targeting DUB1 gene on gastric cancer cell cycle, panel a shows the results of image analysis of MGC803 cells, and panel B shows the detailed quantification results of panel a.

Fig. 7 is a graph showing the results of measuring the effect of siRNA targeting DUB1 gene on the invasion ability of gastric cancer cells, graph a is the result of image analysis of MGC803 cells, graph B is the result of image analysis specifically for graph a, graph C is the result of image analysis for AGS cells, and graph D is the result of image analysis specifically for graph C.

Fig. 8 is a graph showing the results of measuring the effect of siRNA targeting DUB1 gene on the migration ability of gastric cancer cells, graph a is the result of image analysis of MGC803 cells, graph C is the result of detailed quantification of graph a, graph B is the result of image analysis of AGS cells, and graph D is the result of detailed quantification of graph B.

Fig. 9 shows the measurement results of the effect of down-regulated DUB1 gene on the growth of gastric cancer tumor, graph a shows the image analysis results of tumor volume, graph B shows the quantification results of tumor volume at different time after gastric cancer cell injection, and graph C shows the quantification results of tumor weight.

Detailed Description

The invention is described in detail below with reference to the drawings and specific examples, but the invention should not be construed as being limited thereto. The technical means used in the following examples are conventional means well known to those skilled in the art, and materials, reagents and the like used in the following examples can be commercially available unless otherwise specified.

According to the invention, through extensive and intensive research, a DUB (deubiquitinase) siRNA screening library is used for finding that DUB1(GENE ID: 57602; the GENE sequence is shown as SEQ ID NO.1, and the amino acid sequence is shown as SEQ ID NO. 2) is a key regulator for regulating the activity of a Hippo pathway of a gastric cancer patient for the first time, DUB1 is increased in a gastric cancer sample, and poor prognosis of the gastric cancer patient is related to TAZ expression. DUB1 promotes gastric cancer progression through Hippo/TAZ axis, DUB1 is associated with TAZ, inhibits ubiquitination and degradation of k48 linkage, and DUB1 enhances Hippo signaling activity in gastric cancer, and may be a promising marker for diagnosis and treatment of gastric cancer. Additional aspects and advantages of the invention will be set forth in the description which follows of the embodiments, or may be learned by practice of the invention.

Example 1: TCGA data analysis of DUB1 mRNA expression in different stages of gastric cancer

Gene expression data for 385 patients with TCGA gastric cancer were obtained from the GEPIA website (http:// GEPIA. cancer-pku. cn/index. html) and online analysis was performed to compare the expression levels of DUB1 mRNA in normal stomach tissues and different gastric cancer stages; and drawing a box type graph.

The results show that: there was an increase in DUB1 mRNA from different gastric cancer stages compared to normal stomach tissue (fig. 1).

Example 2: tissue chip and immunohistochemical analysis differential expression of DUB1 protein

90 paraffin-embedded gastric cancer samples were obtained from Shanghai super core Biotech (http:// www.superchip.com.cn). The use of this sample was approved by shanghai core biotechnology company and written informed consent was obtained from all patients. The staining density of human samples was detected using specific antibodies against DUB1(Sigma, HPA 12082). The score was calculated based on the intensity and percentage of positive tumor cells in the whole tissue and was scored according to the fromwittz criteria. The staining intensity was graded as: no staining, 0; weak positive, 1; moderate positive, 2, strong positive. The proportion of positive cells is divided into four categories: 0-25% dyeing, 1; 26-50% dyeing, 2; 51-75% staining, 3 and 76-100% staining, 4. Staining 1, 2 low expression, staining 3, 4 high expression. All staining was evaluated at 200 x magnification, and at least 3 fields were calculated per core.

The results show that: DUB1 protein was significantly elevated in gastric cancer samples (fig. 2).

Example 3: detection of interference Effect of DUB1siRNA

1. Cell culture

The gastric cancer cell lines (AGS and MGC803) were cultured in DMEM medium (ThermoFisher Scientific, Cat:11965092) containing 10% fetal bovine serum (Biological Industries, BISH0744), 1% penicillin/streptomycin (DMEM).

2. Design of siRNA

The DUB1siRNA sequence is as follows:

si DUB1#1 sense strand: 3'-CCGGCAAGCUGCGAAUAUUTT-5' as shown in SEQ ID NO. 3;

si DUB1#1 antisense strand: 3'-AAUAUUCGCAGCUUGCCGGT-5' as shown in SEQ ID NO. 4;

si DUB1#2 sense strand: 3'-GCCCACCACUGAAGAGAUUTT-5' as shown in SEQ ID NO. 5;

si DUB1#2 antisense strand: 3'-AAUCUCUUCAGUGGUGUGCTT-5' as shown in SEQ ID NO. 6;

siControl (negative control) sense strand: 3'-UUCUCCGAACGUGUCACGUTT-5' as shown in SEQ ID NO. 7;

siControl (negative control) antisense strand: 3'-ACGUGACACGUUCGGAGAATT-5' as shown in SEQ ID NO. 8.

3. Transfection

Cells were routinely transfected using Lipofectamine 2000 instructions and 48 hours after transfection, knockdown efficiency was measured using RT-qPCR and western blot.

RT-qPCR: total RNA was extracted with the RNeasy Plus Mini Kit (Tiangen, DP451) and reverse transcribed using HiScript II Q RT Supermix (Vazyme, R223-01) according to the manufacturer's specifications. The qRT-PCR used SYBR qPCR Master Mix (Vazyme, Q511-02) and 7500 Rapid real-time PCR System (Singapore applied biosystems), 36B4 as internal controls. The primers used were as follows:

DUB1-F:3'-GAGGCCGGGGCTCTGA-5' as shown in SEQ ID NO. 9;

DUB 1-R: 3'-ACT GGGATGTGCAGACTTGG-5' as shown in SEQ ID NO. 10;

36B 4-F: 3'-GGCGACCTGGAAGACCACCT-5' as shown in SEQ ID NO. 11;

36B 4-R: 3'-CCATCAGCCACCACCTTC-5' as shown in SEQ ID NO. 12.

Western blot: cells were harvested and lysed with RIPA buffer (biomine, # B14001, # B15001) containing a mixture of protease and phosphatase inhibitors. The proteins were electrophoretically separated on SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membrane (Beyotime, # FFN08) using anti-DUB1(Sigma, HPA12082,1: 1000).

The results are shown in fig. 3, and the immunoblot data show that both siRNAs can significantly reduce DUB1 protein levels (fig. 3A and 3B). QPCR assay results showed that DUB1 miRNA could be efficiently silenced by two independent siRNAs (fig. 3C and fig. 3D).

Example 4: effect of knocking down DUB1 on gastric cancer cells and tumors

1. Cell activity assay

MGC803 and AGS cells were transfected with si-DUB 1. 24 hours after transfection, the cell number was collected, and 3000 cells were seeded into a 96-well plate. Cell activity was detected using CCK8 kit (Biomake, # B34302) at the indicated time points.

2. Edu analysis

MGC803 and AGS cells were transfected with siDUB1 or siControl on 24-well plates. 24 hours after transfection, the number of cells was counted, and 4000 cells were seeded into a 96-well plate. Relative cell viability was measured at the indicated time points. The cell number was determined by measuring the absorbance at 450nm using CCK8 cell growth reagent. Cell proliferation was further analyzed by EdU incorporation and flow cytometry. Gastric cancer cells were detected using a 5-ethyl-20-deoxyuridine (EdU) detection kit (Ribobio, Guangzhou, China). For quantitative analysis of the images, each data point represents the positive fluorescence area calculated from at least 5 fields randomly selected from three separate experiments. Incorporation of EdU into FACS assays was performed according to the manufacturer's instructions. These experiments were performed in triplicate.

3. Cell cycle and apoptosis assays

Cell cycle analysis was determined using Propidium Iodide (PI) staining. Cell (1X 10) ⁷ ) Washed with PBS and fixed with 70% ethanol for 30 minutes at room temperature. After three washes with PBS, cells were stained with PI-binding RNaseA (Thermo, # F10797). The red signal was measured with a FACScan (micropore imager). FSC data was analyzed using modfitltv3.1 software. In the apoptosis assay, MGC803 and AGS were double stained with annexin V and Propidium Iodide (PI), using the annexin V-FITC apoptosis detection kit (Vazyme, # A211-02) and Propidium Iodide (PI). Fluorescence was measured with a FACScan (micropore imager). FSC data were analyzed using flowjo7.6 software.

4. Scratch and migration test

In the scratch test, MGC803 and AGS cells containing siDUB1 or siControl were seeded in 6-well plates until confluent and then damaged with sterile tips. Cell pictures were taken at the indicated time points after scraping. The distance between the two sides of the scratch was measured using ImageJ software. Transwell experiments (8 μm pore size, corning) were used for cell migration and invasion, gastric cancer cells migrated to the bottom insert membrane fixed 24 hours after upper chamber coating with matrix (BD bio-coating, usa), stained and counted with crystal violet, and experiments under x 20 objective counted and repeated three times.

5. Clone formation assay

MGC803 and AGS cells containing si-DUB1 or siControl (50nM) were seeded overnight in 6-well plates. After 24 hours, gastric cancer cells were washed with PBS, trypsinized, and plated at low density on six-well plates (approximately 5000 cells per well). Cells were cultured for 10 days, with medium being refreshed every 2 or 3 days. After 10 days, colonies were stained with crystal violet. The number of clones in a given region was calculated.

6. Animal experiments

shDUB1 and shControl were cloned into vector pLKO.1 and entered HEK293 cells with pMD2.G and psPAX2 envelope plasmids. After 48 hours, MGC803 cells were cultured with 200ul of lentivirus in 2ml antibiotic-free medium. The shRNA sequences used were as follows:

shDUB 1: 3'-CCTGTGATTCTCAGGGAACAA-5' as shown in SEQ ID NO. 13;

shControl: 3'-AACAAGATGAAGAGCACCA-5' as shown in SEQ ID NO. 14.

Subcutaneous injection of 2X 10 ⁶ To the lower back area of 4-week-old female BALB/c nude mice, tumor formation was observed for 4 weeks to observe the tumor volume, which was calculated as: tumor volume is 0.5 × square of minimum diameter × maximum diameter, mice are sacrificed 5 weeks after injection, and tumors are removed and weighed.

As a result: CCK8 testing indicated that DUB1 deletion significantly inhibited gastric cancer cell growth (fig. 4). In addition, EdU assay showed that silencing of DUB1 could significantly reduce EdU positive cells in MGC803 and AGS cells (fig. 5). Cell cycle analysis by PI staining and FACS analysis showed that deletion of DUB1 significantly resulted in an increase in the proportion of cells in G1 phase (fig. 6) furthermore, invasion experiments showed that DUB1 is a prerequisite for gastric cancer invasion capacity in AGS and MGC803 cells (fig. 7). Wound healing experiments showed that removal of DUB1 decreased the rate of migration of cancer cells in AGS and MGC803 cells (fig. 8). In vivo tumor growth experiments, stable silencing of DUB1 in MG803 cells could inhibit tumor growth in a transplanted tumor mouse model (fig. 9).

While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.

It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Sequence listing

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<120> DUB1 gene, and application of detection reagent and inhibitor thereof

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ctgcgaattc caggctctaa gaaaagtccc gagggcctca tctccaggac aggctcctcc 1320

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acacccacac acatgccaac catcctagac gaccctggaa agaaggtgaa gaagccagct 1620

cctccacagc acttttcccc cagaactgct caggggctgc ctgggaccag caactcgaat 1680

agcagcagat ctgggagcca aaggcagggc tcctgggaca gcagggatgt tgtcctctct 1740

acctcaccta agctcctggc tacagccact gccaacgggc atgggctgaa ggggaacgac 1800

gagagcgctg gcctcgacag gaggggctcc agcagctcca gcccagagca ctcggccagc 1860

agcgactcca ccaaggcccc ccagaccccc aggagtggag cggcccatct ctgcgattct 1920

caggaaacga actgttccac cgctggccac tccaaaacgc cgccaagtgg agcagattct 1980

aagacggtga agctgaagtc ccctgtcctg agcaacacca ccactgagcc tgcaagcacc 2040

atgtctcctc caccagccaa aaaactggcc ctttctgcca agaaggccag caccctgtgg 2100

agggcgaccg gcaatgacct ccgtccacct cccccctcac catcctccga cctcacccac 2160

cccatgaaaa cctctcaccc cgtcgttgcc tccacttggc ccgtccatag agccagggct 2220

gtgtcacctg ctccccaatc atccagccgc ctgcaacccc ccttcagccc ccaccccaca 2280

ttgctgtcca gtacccccaa gcccccaggg acgtcagaac cacggagctg ctcctccatc 2340

tcgacggcgc tgcctcaggt caacgaggac cttgtgtctc ttccacacca gttgccagag 2400

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cagaggctgg gctcagagac gcgcctccca cagcacatca gggaggccac tgcggctccc 2520

cacgggaaga ggaagaggaa gaagaagaag cgcccggagg acacagctgc cagcgccctg 2580

caggaggggc agacacagag acagcctggg agccccatgt acaggaggga gggccaggca 2640

cagctgcccg ctgtcagacg gcaggaagat ggcacacagc cacaggtgaa tggccagcag 2700

gtgggatgtg ttacggacgg ccaccacgcg agcagcagga agcggaggag gaaaggagca 2760

gaaggtcttg gtgaagaagg cggcctgcac caggacccac ttcggcacag ctgctctccc 2820

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agaaagcagg agacacagcg ggcagtagaa gaggatgggc atctcaaatg cccaaggagt 2940

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Lys Lys Arg Pro Glu Asp Thr Ala Ala Ser Ala Leu Gln Glu Gly Gln

850 855 860

Thr Gln Arg Gln Pro Gly Ser Pro Met Tyr Arg Arg Glu Gly Gln Ala

865 870 875 880

Gln Leu Pro Ala Val Arg Arg Gln Glu Asp Gly Thr Gln Pro Gln Val

885 890 895

Asn Gly Gln Gln Val Gly Cys Val Thr Asp Gly His His Ala Ser Ser

900 905 910

Arg Lys Arg Arg Arg Lys Gly Ala Glu Gly Leu Gly Glu Glu Gly Gly

915 920 925

Leu His Gln Asp Pro Leu Arg His Ser Cys Ser Pro Met Gly Asp Gly

930 935 940

Asp Pro Glu Ala Met Glu Glu Ser Pro Arg Lys Lys Lys Lys Lys Lys

945 950 955 960

Arg Lys Gln Glu Thr Gln Arg Ala Val Glu Glu Asp Gly His Leu Lys

965 970 975

Cys Pro Arg Ser Ala Lys Pro Gln Asp Ala Val Val Pro Glu Ser Ser

980 985 990

Ser Cys Ala Pro Ser Ala Asn Gly Trp Cys Pro Gly Asp Arg Met Gly

995 1000 1005

Leu Ser Gln Ala Pro Pro Val Ser Trp Asn Gly Glu Arg Glu Ser Asp

1010 1015 1020

Val Val Gln Glu Leu Leu Lys Tyr Ser Ser Asp Lys Ala Tyr Gly Arg

1025 1030 1035 1040

Lys Val Leu Thr Trp Asp Gly Lys Met Ser Ala Val Ser Gln Asp Ala

1045 1050 1055

Ile Glu Asp Ser Arg Gln Ala Arg Thr Glu Thr Val Val Asp Asp Trp

1060 1065 1070

Asp Glu Glu Phe Asp Arg Gly Lys Glu Lys Lys Ile Lys Lys Phe Lys

1075 1080 1085

Arg Glu Lys Arg Arg Asn Phe Asn Ala Phe Gln Lys Leu Gln Thr Arg

1090 1095 1100

Arg Asn Phe Trp Ser Val Thr His Pro Ala Lys Ala Ala Ser Leu Ser

1105 1110 1115 1120

Tyr Arg Arg

<210> 3

<211> 19

<212> DNA/RNA

<213> Artificial sequence

<400> 3

ccggcaagcu gcgaauauu 19

<210> 4

<211> 20

<212> DNA/RNA

<213> Artificial sequence

<400> 4

aauauucgca gcuugccggt 20

<210> 5

<211> 21

<212> DNA/RNA

<213> Artificial sequence

<400> 5

gcccaccacu gaagagauut t 21

<210> 6

<211> 21

<212> DNA/RNA

<213> Artificial sequence

<400> 6

aaucucuuca guggugugct t 21

<210> 7

<211> 21

<212> DNA/RNA

<213> Artificial sequence

<400> 7

uucuccgaac gugucacgut t 21

<210> 8

<211> 21

<212> DNA/RNA

<213> Artificial sequence

<400> 8

acgugacacg uucggagaat t 21

<210> 9

<211> 16

<212> DNA

<213> Artificial sequence

<400> 9

gaggccgggg ctctga 16

<210> 10

<211> 20

<212> DNA

<213> Artificial sequence

<400> 10

actgggatgt gcagacttgg 20

<210> 11

<211> 20

<212> DNA

<213> Artificial sequence

<400> 11

ggcgacctgg aagaccacct 20

<210> 12

<211> 18

<212> DNA

<213> Artificial sequence

<400> 12

ccatcagcca ccaccttc 18

<210> 13

<211> 21

<212> DNA/RNA

<213> Artificial sequence

<400> 13

cctgtgattc tcagggaaca a 21

<210> 14

<211> 19

<212> DNA/RNA

<213> Artificial sequence

<400> 14

aacaagatga agagcacca 19

Claims (10)

1. Application of a reagent for detecting the expression level of DUB1 gene in preparing a product for diagnosing gastric cancer.

2. The use of claim 1, wherein the DUB1 gene is expressed in elevated levels in a gastric cancer patient.

3. The use according to claim 1, wherein the agent is selected from the group consisting of:

a probe that specifically recognizes the DUB1 gene; or

Primers that specifically amplify the DUB1 gene; or

A specific binding agent that specifically binds to a protein encoded by the DUB1 gene.

4. The use according to claim 3, wherein the primer sequence for specifically amplifying the DUB1 gene is shown in SEQ ID No. 9-10.

5. Use of the DUB1 gene in screening for potential substances for treating gastric cancer according to claim 1, wherein the substances are substances that down-regulate the expression level of the DUB1 gene.

6. Use of the inhibitor of the expression of DUB1 gene of claim 1 in the manufacture of a medicament for the treatment of gastric cancer.

7. The use of claim 6, wherein the inhibitor comprises an siRNA capable of targeting interference with the DUB1 gene.

8. The use of claim 7, wherein the siRNA has the sequence shown in SEQ ID No. 3-6.

9. A pharmaceutical composition for treating gastric cancer, comprising a therapeutically effective amount of the inhibitor of claim 6.

10. The pharmaceutical composition of claim 9, further comprising other drugs compatible with the inhibitor and a pharmaceutically acceptable carrier and/or adjuvant.

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